CD62p 阳性率是保存血小板质量监测的重要指标。为在静脉全血内血小板CD62p 测定方法的基础上优化建立流式细胞术(FCM) 测定保存血小板表达CD62p 的方法在血小板检测标本内加入0. 1 mmol/ L 潘生丁稳定血小板;对TBS 溶液进行了改良添加了1. 1 mmol/ L 的EDTA 和100μmol/ L 潘生丁用以代替PBS;采用新鲜富含血小板血浆( FPRP) 制备的阴、阳性对照;进行方法学评价。

关键词：血小板流式细胞术CD62p

欧阳锡林刘景汉石　群高大勇1 2
(医院输血科临床输血中心北京100853 ;1 科技大学热科学及能源工程系
低温生物医学研究所合肥230027 ; 2Department of Mechanical Engineering University of
Kentucky Lexington KY 40506 - 0108 USA)
摘要：CD62p 阳性率是保存血小板质量监测的重要指标。为在静脉全血内血小板CD62p 测定方法的基础上优化建立流式细胞术(FCM) 测定保存血小板表达CD62p 的方法在血小板检测标本内加入0. 1 mmol/ L 潘生丁稳定血小板;对TBS 溶液进行了改良添加了1. 1 mmol/ L 的EDTA 和100μmol/ L 潘生丁用以代替PBS;采用新鲜富含血小板血浆( FPRP) 制备的阴、阳性对照;进行方法学评价。结果表明:加入0. 1 mmol/ L 潘生丁和1. 1 mmol/ L 的EDTA 可以有效防止实验过程中的血小板人为激活;采用新鲜富含血小板血浆( FPRP) 制备的阴、阳性对照既可优化FCM 分析方法还可用于检验所购荧光抗体的质量保证实验结果的有效性。采用泛血小板特异性标志物CD61 可灵敏特异地识别血小板提高了实验抗干扰能力。制备保存血小板时采用的是大号采血针抽血流畅对血小板CD62p 表达测定人为影响很小明显优于用注射器采集患者静脉全血的做法。CD62p 测定灵敏度可达1 % 制备的标本可稳定48 小时。结论:利用该流式细胞术可以灵敏而特异地测定保存血小板的CD62P 表达率;该方法简便易行提高了结果的准确性具有良好的稳定性和重复性。
关键词：流式细胞术; 血小板保存; CD62p
Modif ied Flow Cytometry Assay for CD62p Expression of Preserved
Platelets
OU YAN G Xi2Lin L IU J ing2Han SHI Qun GAO Da2Yong1 2
( Department of Blood Transf usion General Hospital of PL A Center of Cli nical Transf usion of PL A Beiji ng 100853 Chi na ;
1 Instit ute of Cryobiomedici ne Faculty of Thermal Science and Energy Engi neeri ng University of Sciences and Technologies of Chi na
Hef ei 230027 Chi na ; 2 Department of Mechanical Engi neeri ng University of Kent ucky Lexi ngton KY 40506 - 0108 USA )
itive platelets is simple and efficient .
Abstract ：CD62p was an important monitoring parameter for preserved platelets quality. To set up an optimized flow cytometry assay for preserved platelets d on the CD62p on platelets the platelet samples were collected 0. 1 mmol/ L persantine and 1. 1 mmol/ L EDTA were added into the modified TBS used to replace PBS dilution ; the methodological evaluation were carried out . Results showed that 0. 1 mmol/ L persantine and 1. 1 mmol/ L EDTA achieved to prevent platelets activation during the test procedure. The favorable negative or positive samples were prepared for check of fluorescence antibody′s quality to ensure the validity of results. CD61 was used to identify platelets for assay and improve veracity of assay. The special injector was also replaced by special big syringe needle for blood collection to reduce in vit ro artifacts and the prepared sample can be steady2going for 48 hours at 4 ℃after fixed by 1 % paraformaldehyde. It is concluded that this flow cytometry assay for CD62p pos
Key words ：flow cytometry ; platelet preservation ; CD62p